| Assay Method Information | |
| | Inhibitory Assay |
| Description: | The IC50 values for the tested compounds were determined as previously described [2, 3], with minor modifications. Briefly, in 96-well tissue culture plates, 2.4x104 Vero-E6 cells were distributed in each well and incubated overnight at a humidified 37° C. incubator under 5% CO2 condition. The cell monolayers were then washed once with 1xPBS. An aliquot of the SARS-CoV-2 "NRC-03-nhCoV" virus [40] containing 100 TCID50 was incubated with serial diluted compounds and kept at 37° C. for 1 h. The Vero-E6 cells were treated with virus/compound mix and co-incubated at 37° C. in a total volume of 200 uL per well. Untreated cells infected with virus represented the virus control; however, cells that were not treated and not infected were the cell control. Following incubation at 37° C. in 5% CO2 incubator for 72 h, the cells were fixed with 100 uL of 10% paraformaldehyde for 20 min and stained with 0.5% crystal violet in distilled water for 15 min at RT. The crystal violet dye was then dissolved using 100 uL of absolute methanol per well and the optical density of the color was measured at 570 nm using the Anthos Zenyth 200 rt plate reader (Anthos Labtec Instruments, Heerhugowaard, the Netherlands). The IC50 of the compound refers to a concentration of the compound required to reduce the virus-induced cytopathic effect (CPE) by 50%, relative to the virus control. |
| Affinity data for this assay | |
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